• Title/Summary/Keyword: isolation and purification

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Isolation of a Calcium-binding Peptide from Chlorella Protein Hydrolysates

  • Jeon, So-Jeong;Lee, Ji-Hye;Song, Kyung-Bin
    • Preventive Nutrition and Food Science
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    • v.15 no.4
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    • pp.282-286
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    • 2010
  • To isolate a calcium-binding peptide from chlorella protein hydrolysates, chlorella protein was extracted and hydrolyzed using Flavourzyme, a commercial protease. The degree of hydrolysis and calcium-binding capacity were determined using trinitrobenzenesulfonic acid and orthophenanthroline methods, respectively. The enzymatic hydrolysis of chlorella protein for 6 hr was sufficient for the preparation of chlorella protein hydrolysates. The hydrolysates of chlorella protein were then ultra-filtered under 5 kDa as molecular weight. The membrane-filtered solution was fractionated using ion exchange, reverse phase, normal phase chromatography, and fast protein liquid chromatography to identify a calcium-binding peptide. The purified calcium-binding peptide had a calcium binding activity of 0.166 mM and was determined to be 700.48 Da as molecular weight, and partially identified as a peptide containing Asn-Ser-Gly-Cys based on liquid chromatography/electrospray ionization tandem mass spectrum.

Role of Unstable Phenanthrene-Degrading Pseudomonas species in Natural Attenuation of Phenanthrene-Contaminated Site

  • Prakash, Om;Lal, Rup
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.79-87
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    • 2013
  • An unstable yet efficient phenanthrene-degrading bacterium strain Ph-3 was isolated from a petroleum-contaminated site at the Mathura Oil Refinery, India. The strain was identified as Pseudomonas sp. using a polyphasic approach. An analysis of the intermediates and assays of the degradative enzymes from a crude extract of phenanthrene-grown cells showed a novel and previously unreported pattern of 1, 2-dihydroxy naphthalene and salicylic acid production. While strain Ph-3 lost its phenanthrene- degrading potential during successive transfers on a rich medium, it maintained this trait in oligotrophic soil conditions under the stress of the pollutant and degraded phenanthrene efficiently in soil microcosms. Although the maintenance and in vitro study of unstable phenotypes are difficult and such strains are often missed during isolation, purification, and screening, these bacteria constitute a substantial fraction of the microbial community at contaminated sites and play an important role in pollutant degradation during biostimulation or monitored natural attenuation.

Isolation of the Arabidopsis Phosphoproteome Using a Biotin-tagging Approach

  • Kwon, Sun Jae;Choi, Eun Young;Seo, Jong Bok;Park, Ohkmae K.
    • Molecules and Cells
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    • v.24 no.2
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    • pp.268-275
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    • 2007
  • Protein phosphorylation plays a key role in signal transduction in cells. Since phosphoproteins are present in low abundance, enrichment methods are required for their purification and analysis. Chemical derivatization strategies have been devised for enriching phosphoproteins and phosphopeptides. In this report, we employed a strategy that replaces the phosphate moieties on serine and threonine residues with a biotin-containing tag via a series of chemical reactions. Ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBISCO)-depleted protein extracts prepared from Arabidopsis seedlings were chemically modified for 'biotin-tagging'. The biotinylated (previously phosphorylated) proteins were then selectively isolated by avidin-biotin affinity chromatography, followed by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). This led to the identification of 31 protein spots, representing 18 different proteins, which are implicated in a variety of cellular processes. Despite its current technical limitations, with further improvements in tools and techniques this strategy may be developed into a useful approach.

Isolation, purification and characterization of phytohemagglutinating proteins from Korean natural products

  • Chung, See-Ryun;Jeune-Chung, Kyung-Hee;Kim, Kyong-Ae
    • Archives of Pharmacal Research
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    • v.3 no.1
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    • pp.31-36
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    • 1980
  • Seeds or beans of 55 plants belonging to 31 families were screened by using several different types of red blood cells to find new lectins. In this paper, white kidney been (phaseolus vulgaris C.) was chosen to study biochemical properties of hemagglutinating proteins(lectins). An anion exchanger, DEAE Sephadex A-50, and polyacrylamide disc gel electrophoresis were main techniques used. From three main fractions eluted by stepwise NaCl gradient in 25mM Tris-HCI buffer on DEAE Sephadex column, principal lectin was identified.

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Laboratory Experiment: Synthesis and Characterization of 4-Methyl-N-(phenylacetyl)benzenesulfonamide through Cu(I)-Catalysis

  • Jung, Byunghyuck
    • Journal of the Korean Chemical Society
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    • v.62 no.3
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    • pp.187-190
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    • 2018
  • A three-component coupling reaction of phenylacetylene, p-toluenesulfonyl azide, and water through copper catalysis is described to provide knowledge of spectroscopy and catalytic reactions and to introduce current research topics in organic chemistry for second-year undergraduate students. In the presence of stoichiometric amounts of phenylacetylene, p-toluenesulfonyl azide, and triethylamine, the reaction was performed with 4 mol% CuCl in water as the sole solvent and was completed in 1.5 h. A practical purification method and recrystallization of the crude reaction mixture resulted in the rapid isolation of the desired product with yields of 42~65%. Students characterized 4-methyl-N-(phenylacetyl)benzenesulfonamide by using melting-point determination, infrared spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. This experimental procedure and spectroscopic data analysis will serve as a platform for students to apply classroom knowledge in practical state-of-the-art research.

Radical Scavenging Activity of Gallic Acid from Woodfordia Fruticosa Flowers

  • Bhatt, Lok Ranjan;Yook, Chan-Nam;Choi, Hwa-Jung;Baek, Seung-Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.903-906
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    • 2008
  • Woodfordia fruticosa Kurz (Lythraceae) is used in the treatment of various ailments in traditional medicines. DPPH activity guided fractionation and purification process was used to identify the free radical-scavenging components from the flowers of this plant. The methanolic extract of the plant was first fractionated into four extracts; namely, n-hexane, chloroform, ethyl acetate and water fractions. Among them, the ethyl acetate fraction was found to be the most effective and was further subjected to activity guided-fractionation and isolation procedures. After successive column chromatography on silica gel and Sephadex LH-20, gallic acid, which is responsible for the radical scavenging activity, was isolated and its structure was elucidated by spectral methods ($^1H$ NMR, $^{13}C$ NMR) and by comparison with literature.

Screening of Antifungal Compounds from Microorganisms with Preferential Activity against the Mycelial Phase of Candida albicans (토양으로부터 Candida albicans 의 균사형태에 선택적인 활성을 나타내는 미생물로부터 항진균 물질의 탐색)

  • Kim, Sung-Uk;Nam, Ji-Youn;Kwon, Byoung-Mok;Son, Kwang-Hee;Bok, Song Hae
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.170-177
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    • 1995
  • To search and develop the compounds exhibiting antifungal activities against the mycelial phase of Candida albicans, approximately 2,900 microorganisms isolated from soil were examined for antifungal activity. Among them, a strain with preferential activity against the mycelial phase of Candida albicans was isolated and identified as Streptomyces sp. A393. Isolation and purification of compounds A393 showing antifungal activity against the mycelial phase of C. albicans were performed using XAD-7 column chromatography, silica gel chromatography, preparative thin- layer silica gel chromatography, and HPLC. The molecular weights of compounds isolated from Streptomyces sp. A393 were determined as 774, 790, 804 and 820. These compounds appeared to have a structure of macrolide antibiotics, oligomycin A, B, C and E. Especially, oligomycin E, which is formerly reported to have no antifungal activity, showed antifungal activity against the mycelial phase of Candida albicans.

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Adding Effect of TCA-Precipitated Blood Powder to the Phenol Formaldehyde Resin for Plywood (합판용 페놀수지 접착제에 대한 TCA침전 혈액분말의 첨가효과)

  • Lee, Hwa-Hyoung;Lee, Jong-Shin;Jang, Sang-Sik
    • Journal of the Korean Wood Science and Technology
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    • v.24 no.2
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    • pp.15-19
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    • 1996
  • To utilize the waste materials and develope wood adhesive from isolated bloods of slaughtered cow and pig and also to prevent water pollution, simple and rapid method of isolation and purification of plasma proteins from pig bloods with trichloroacetic acid(TCA) treatment was developed. Adding of TCA-precipitated blood powder to the phenol formaldehyde resin(PF) improved dry and wet strength of plywood and resulted in fast hot pressing times.

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Isolation and Purification of Acidic Polysaccharide of Korean Red Ginseng Acting against Toxic Action of Toxohormone-L from Cancerous Ascites Fluid (암독소(Toxohormone-L)의 작용을 저해하는 홍삼산성다당체의 분리 및 정제)

  • Lee, Seong-Dong;Hwang, Yun-Gyeong
    • The Korean Journal of Food And Nutrition
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    • v.3 no.2
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    • pp.133-140
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    • 1990
  • Toxohormone-L is a lipolytic factor, found in ascites fluid of sarcoma 180-bearing mice and of patients with hepatoma A substance that inhibited the lipolytic action of Toxohormoue-L was isolated and purified from Korean red ginseng powder. This substance had a pectin-like $\alpha$-1,4-polygalacturonan backbone with some acetoxyl groups, and so was an acidic polysaccharide It inhibited Toxohormone-L Induced liploysis in a dose dependent manner at concentrations higher than 10 Ug/ml. Purified acidic Polysaccharide yield(PG4-3 and PG4-4 fraction) was about 0.03%. And also pectic acid that inhibited the lipolytic action of Toxohormone-L.

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